Biochemical exploration of family GH119 reveals a single α-amylase specificity and confirms shared catalytic machinery with GH57 enzymes.

While hundreds of starch- and glycogen-degrading enzymes have been characterized experimentally in historical families such as GH13, GH14, GH15, GH57 and GH126 of the CAZy database (www.cazy.org), the α-amylase from Bacillus circulans is the only enzyme that has been characterized in family GH119. Since glycosidase families have been shown to often group enzymes with different substrates or products, a single characterized enzyme in a family is insufficient to extrapolate enzyme function based solely on sequence similarity. Here we report the rational exploration of family GH119 through the biochemical characterization of five GH119 members. All enzymes shared single α-amylase specificity but display distinct product profile. We also report the first kinetic constants in family GH119 and the first experimental validation of previously predicted catalytic residues in family GH119, confirming that families GH119 and GH57 can be grouped in the novel clan GH-T of the CAZy database.

Investigation and drug design for novel molecules from natural products as inhibitors for controlling multiple myeloma disease using in-silico tools.

Multiple myeloma (MM) is a disease that causes plasma cell growth in the bone marrow and immune globulin buildup in blood and urine. Despite recent advances in MM therapy, many still die due to its high mortality rate. A study using computational simulations analyzed 100 natural ingredients from the SANC database to determine if they inhibited the IgH domain, a known cause of multiple myeloma. Natural component Diospyrin inhibited the IgH enzyme with the best binding energy of -10.3 kcal/mol and three carbon-hydrogen bonds, followed by Parviflorone F complex with a binding energy of -10.1 kcal/mol and two conventional-hydrogen bonds. As a result, the Molecular Dynamic simulation was used to test the stability of the two complexes. During the simulation, the Diospyrin molecule dissociated from the protein at roughly 67.5 ns, whereas the Parviflorone F molecule stayed attached to the protein throughout. The latter was the subject of the investigation. The analysis of the production run data revealed that the Parviflorone F molecule exhibits a variety of conformations within the binding pocket while keeping a relatively constant distance from the protein's center of mass. The analysis of the production run data revealed that the Parviflorone F molecule exhibited a variety of conformations within the binding pocket while keeping a relatively constant distance from the protein's center of mass. The root mean square deviation (RMSD) plots for both the protein and complex showed a stable and steady average value of 4.4 Å for the first 82 nanoseconds of manufacture. As a result, the average value increased to 8.3 Å. Furthermore, the components of the binding free energy, as computed by MM-GBSA, revealed that the mean binding energy of the Parviflorone F molecule was -23.88 kcal/mol. Finally, after analyzing all of the examination data, Parviflorone F was identified as a powerful inhibitor of the IgH domain and hence of the MM disease, which requires further in-vivo conformation.Communicated by Ramaswamy H. Sarma.

Perceptions and Attitudes of Primary Care Physicians Toward Dermatologic Surgery.

Purpose: Dermatologic surgery is a well-established subspecialty in dermatology that involves various therapeutic and esthetic procedures. To date, perceptions of primary care physicians (PCPs) regarding dermatologic surgery are poorly understood. This study aimed to assess the perception and attitude of PCPs toward dermatologic surgery and explore the factors that might affect their perception and attitude toward it. Patients and Methods: This cross-sectional study used an online survey. The survey was distributed among PCPs working in various regions of Saudi Arabia and contained questions about demographics, medical training information, participants' awareness of dermatologic surgery, and a list of 10 cutaneous procedures to select the most qualified physician performing these procedures. Univariate and multivariate analyses were performed. P≤0.05 was considered statistically significant. Results: In total, 440 PCPs participated in this study. Overall, 70% of the PCPs had heard about dermatologic surgery, whereas 30% had never heard about it. PCPs reported that dermatologic surgeons were the most qualified physicians to perform laser procedures (60.5%), hair transplantation (60.5%), excision of small benign and malignant skin tumors (46.1%), excision of small melanocytic nevi (55.7%), filler injection (44.8%), and injection of botulinum toxin (48.9%); 64.5% considered plastic surgeons to be the most qualified for liposuction. Both physicians were equally selected for performing cyst excision, procedures involving the fingernails and toenails, and scar correction. Multivariate analysis showed that female physicians had heard significantly more about dermatologic surgery (P=0.047, OR: 1.57, 95% CI: 1.006-2.45) and reported that dermatologic surgeons were the most qualified physicians (P=0.042, OR: 1.51, 95% CI: 1.016-2.25) than male physicians. PCPs working at secondary and tertiary health care hospitals heard more about dermatologic surgery (P=0.015, OR: 2.36, 95% CI: 1.18-4.71) than those working at primary health care centers. Conclusion: Most PCPs were aware of dermatologic surgery and recognized dermatologic surgeons as the most qualified physicians to perform most cutaneous procedures.

Computational and experimental analysis of foxtail millet under salt stress and selenium supplementation.

Salinity stress is a major threat to crop growth and productivity. Millets are stress-tolerant crops that can withstand the environmental constraints. Foxtail millet is widely recognized as a drought and salinity-tolerant crop owing to its efficient ROS scavenging mechanism. Ascorbate peroxidase (APX) is one of the reactive oxygen species (ROS) scavenging enzymes that leads to hydrogen peroxide (H2O2) detoxification and stabilization of the internal biochemical state of the cell under stress. This inherent capacity of the APX enzyme can further be enhanced by the application of an external mitigant. This study focuses on the impact of salt (NaCl) and selenium (Se) application on the APX enzyme activity of foxtail millet using in silico and in-vitro techniques and mRNA expression studies. The NaCl was applied in the concentrations, i.e., 150 mM and 200 mM, while the Se was applied in 1 µM, 5 µM, and 10 µM concentrations. The in silico studies involved three-dimensional structure modeling and molecular docking. The in vitro studies comprised the morphological and biochemical parameters, alongside mRNA expression studies in foxtail millet under NaCl stress and Se applications. The in silico studies revealed that the APX enzyme showed better interaction with Se as compared to NaCl, thus suggesting the enzyme-modulating role of Se. The morphological and biochemical analysis indicated that Se alleviated the NaCl (150 mM and 200 mM) and induced symptoms at 1 µM as compared to 5 and 10 µM by enhancing the morphological parameters, upregulating the gene expression and enzyme activity of APX, and ultimately reducing the H2O2 content significantly. The transcriptomic studies confirmed the upregulation of chloroplastic APX in response to salt stress and selenium supplementation. Hence, it can be concluded that Se as a mitigant at lower concentrations can alleviate NaCl stress in foxtail millet.

Exogenous zinc mitigates salinity stress by stimulating proline metabolism in proso millet (Panicum miliaceum L.).

Salinity is one of the most concerning ecological restrictions influencing plant growth, which poses a devastating threat to global agriculture. Surplus quantities of ROS generated under stress conditions have negative effects on plants' growth and survival by damaging cellular components, including nucleic acids, lipids, proteins and carbohydrates. However, low levels of ROS are also necessary because of their role as signalling molecules in various development-related pathways. Plants possess sophisticated antioxidant systems for scavenging as well as regulating ROS levels to protect cells from damage. Proline is one such crucial non-enzymatic osmolyte of antioxidant machinery that functions in the reduction of stress. There has been extensive research on improving the tolerance, effectiveness, and protection of plants against stress, and to date, various substances have been used to mitigate the adverse effects of salt. In the present study Zinc (Zn) was applied to elucidate its effect on proline metabolism and stress-responsive mechanisms in proso millet. The results of our study indicate the negative impact on growth and development with increasing treatments of NaCl. However, the low doses of exogenous Zn proved beneficial in mitigating the effects of NaCl by improving morphological and biochemical features. In salt-treated plants, the low doses of Zn (1 mg/L, 2 mg/L) rescued the negative impact of salt (150mM) as evidenced by increase in shoot length (SL) by 7.26% and 25.5%, root length (RL) by 21.84% and 39.07% and membrane stability index (MSI) by 132.57% and 151.58% respectively.The proline content improved at all concentrations with maximum increase of 66.65% at 2 mg/L Zn. Similarly, the low doses of Zn also rescued the salt induced stress at 200mM NaCl. The enzymes related to proline biosynthesis were also improved at lower doses of Zn. In salt treated plants (150mM), Zn (1 mg/L, 2 mg/L) increased the activity of P5CS by 19.344% and 21%. The P5CR and OAT activities were also improved with maximum increase of 21.66% and 21.84% at 2 mg/L Zn respectively. Similarly, the low doses of Zn also increased the activities of P5CS, P5CR and OAT at 200mM NaCl. Whereas P5CDH enzyme activity showed a decrease of 82.5% at 2mg/L Zn+150mM NaCl and 56.7% at 2mg/L Zn+200 mM NaCl. These results strongly imply the modulatory role of Zn in maintaining of proline pool during NaCl stress.

Oral Bioactive Self-Nanoemulsifying Drug Delivery Systems of Remdesivir and Baricitinib: A Paradigmatic Case of Drug Repositioning for Cancer Management.

Oral anticancer therapy mostly faces the challenges of low aqueous solubility, poor and irregular absorption from the gastrointestinal tract, food-influenced absorption, high first-pass metabolism, non-targeted delivery, and severe systemic and local adverse effects. Interest has been growing in bioactive self-nanoemulsifying drug delivery systems (bio-SNEDDSs) using lipid-based excipients within nanomedicine. This study aimed to develop novel bio-SNEDDS to deliver antiviral remdesivir and baricitinib for the treatment of breast and lung cancers. Pure natural oils used in bio-SNEDDS were analyzed using GC-MS to examine bioactive constituents. The initial evaluation of bio-SNEDDSs were performed based on self-emulsification assessment, particle size analysis, zeta potential, viscosity measurement, and transmission electron microscopy (TEM). The single and combined anticancer effects of remdesivir and baricitinib in different bio-SNEDDS formulations were investigated in MDA-MB-231 (breast cancer) and A549 (lung cancer) cell lines. The results from the GC-MS analysis of bioactive oils BSO and FSO showed pharmacologically active constituents, such as thymoquinone, isoborneol, paeonol and p-cymenene, and squalene, respectively. The representative F5 bio-SNEDDSs showed relatively uniform, nanosized (247 nm) droplet along with acceptable zeta potential values (+29 mV). The viscosity of the F5 bio-SNEDDS was recorded within 0.69 Cp. The TEM suggested uniform spherical droplets upon aqueous dispersions. Drug-free, remdesivir and baricitinib-loaded bio-SNEDDSs (combined) showed superior anticancer effects with IC50 value that ranged from 1.9-4.2 µg/mL (for breast cancer), 2.4-5.8 µg/mL (for lung cancer), and 3.05-5.44 µg/mL (human fibroblasts cell line). In conclusion, the representative F5 bio-SNEDDS could be a promising candidate for improving the anticancer effect of remdesivir and baricitinib along with their existing antiviral performance in combined dosage form.

Selenate and selenite transporters in proso millet: Genome extensive detection and expression studies under salt stress and selenium.

Crops are susceptible to a variety of stresses and amongst them salinity of soil is a global agronomic challenge that has a detrimental influence on crop yields, thus posing a severe danger to our food security. Therefore, it becomes imperative to examine how plants respond to salt stress, develop a tolerance that allows them to live through higher salt concentrations and choose species that can endure salt stress. From the perspective of food, security millets can be substituted to avoid hardships because of their efficiency in dealing with salt stress. Besides, this problem can also be tackled by using beneficial exogenous elements. Selenium (Se) which exists as selenate or selenite is one such cardinal element that has been reported to alleviate salt stress. The present study aimed for identification of selenate and selenite transporters in proso millet (Panicum miliaceum L.), their expression under NaCl (salt stress) and Na2SeO3 (sodium selenite)treatments. This study identified eight transporters (RLM65282.1, RLN42222.1, RLN18407.1, RLM74477.1, RLN41904.1, RLN17428.1, RLN17268.1, RLM65753.1) that have a potential role in Se uptake in proso millet. We analyzed physicochemical properties, conserved structures, sub-cellular locations, chromosome location, molecular phylogenetic analysis, promoter regions prediction, protein-protein interactions, three-dimensional structure modeling and evaluation of these transporters. The analysis revealed the chromosome location and the number of amino acids present in these transporters as RLM65282.1 (16/646); RLN42222.1 (1/543); RLN18407.1 (2/483); RLM74477.1 (15/474); RLN41904.1 (1/521); RLN17428.1 (2/522); RLN17268.1(2/537);RLM65753.1 (16/539). The sub-cellular locations revealed that all the selenite transporters are located in plasma membrane whereas among selenate transporters RLM65282.1 and RLM74477.1 are located in mitochondria and RLN42222.1 and RLN18407.1 in chloroplast. The transcriptomic studies revealed that NaCl stress decreased the expression of both selenate and selenite transporters in proso millet and the applications of exogenous 1µM Se (Na2SeO3) increased the expression of these Se transporter genes. It was also revealed that selenate shows similar behavior as sulfate, while selenite transport resembles phosphate. Thus, it can be concluded that phosphate and sulphate transporters in millets are responsible for Se uptake.

Evaluation of Scalp Hair Density and Diameter in the Arab Population: Clinical Office-Based Phototrichogram Analysis.

Background: Knowing normal values of scalp hair thickness and density is beneficial for diagnosing hair disorders, monitoring responses to therapy, performing hair transplants, and conducting relevant research. Such normal values have not yet been studied for Arabs. Objective: To assess the hair density and diameter of the Arab population at different scalp sites using phototrichogram. Methods: A cross-sectional descriptive study was performed at King Saud University Medical City from December 2021 to August 2022. The study was conducted on Arab adults who were not suffering from any hair disorder. The study was conducted using a questionnaire and clinical examination by phototrichogram. Results: A total of 120 participants were enrolled in this study. The mean hair density was 147.1± 7.8 hairs/cm2, and the mean hair diameter was 87±4.9 µm. There were significant differences between hair density (P=0.0001) and diameter (P=0.0001) in different regions of the scalp. Females had higher hair density than males at occiput (P=0.038). Conclusion: Arab hair has a higher thickness and lower density than hair of other ethnicities. This needs to be considered when diagnosing and/or treating hair problems in Arabs.

Apoptosis-mediated anti-proliferative activity of Calligonum comosum against human breast cancer cells, and molecular docking of its major polyphenolics to Caspase-3.

Due to poor diagnosis breast cancer in women has emerged as the most common cause of death disease in developing countries. Medicinal plants have been used for thousands of years and can be useful in healthcare, especially in developing countries. Ethanol extracts of leaves of fire bush or arta (Calligonum comosum; EECC), exhibited significant anticancer potencies against two breast cancer cell lines, MCF-7 and MDA 231. These in vitro effects of EECC indicated potential anticancer activities that were determined to be specific since minimal toxicity was recorded against MCF-12, a non-cancerous breast cell line used as a reference. EECC also induced cell cycle arrest in MCF-7 and MDA 231 as revealed by the increased proportions of sub-G1 cells. Fluorescence-activated cell sorter analysis (FACS), utilizing double staining by annexin V-FITC/propidium iodide, revealed that the observed cytotoxic effects were mediated via apoptosis and necrosis. FACS measurement of thegreater in fluorescence intensity, linked with oxidation of DCFH to DCF, revealed that apoptosis was attributable to production of free radicals. EECC-mediated apoptosis was further validated by observation of up-regulation in the "executioner" enzyme, caspase 3. The current findings reveal that EECC exhibits significant, selective cytotoxicity to breast cancer cells, that proceeds via the generation of ROS, which culminates in apoptosis. The anti-proliferative effects of EECC weres further verified by use of a structure-based, virtual screening between its major bioactive polyphenolic constituents and the apoptosis executioner marker enzyme, caspase-3. Based on their glide score values against the active site of caspase 3, some phyto-constituents present in EECC, such as DL-alpha-tocopherol and campesterol, exhibited distinctive, drug-like potential with no predicted toxicity to non-target cells. Taken together, the usefulness of natural phenolic and flavonoid compounds contained in Calligonum comosum were suggested to be potent anticancer agents.

Effects of blood meal sources on the biological characteristics of Aedes aegypti and Culex pipiens (Diptera: Culicidae).

Mosquitoes transmit many diseases to humans and animals e.g., malaria, yellow fever, dengue, filariasis and encephalitis. The fundamental target of this search was to study the effect of three different blood meal sources (human; rabbit and pigeon) on some biological and behavioral properties of Aedes aegypti and Culex pipiens. The results have assured that the females of the mosquito Ae. aegypti that were fed on human blood meal has registered the highest feeding activity from feeding on the blood meal whereas the females of the other mosquito Cx. pipiens have shown the highest feeding activity after being fed on pigeons when compared with its feeding on other factors. The results have shown non-significant variation in the average time necessary to digest the blood meal on both mosquito species Ae. aegypti and Cx. pipiens that were fed on vertebrate hosts under laboratory conditions. Furthermore, results assured that the difference in blood meal sources has yielded distinct variation in the reproductive capacity and efficiency of both female mosquitoes under investigation where both species Ae. aegypti and Cx. pipiens already fed on human blood meal have yielded a pronounced distinctive increase in egg production (oviposition) when compared with females that were fed on pigeon or rabbit blood meal respectively. Moreover, feeding of the female mosquitoes under lab conditions on different blood meal sources did not affect the level of the hatching eggs that were laid by both mosquito females.

Seagrasses extracts as potential mosquito larvicides in Saudi Arabia.

The present study aimed to evaluate the toxic and biological effects of some extracts of seagrasses (Cymodocea rotundata; Halophila ovata& Thalassia hemprichii) against Aedes aegypti, which transmits dengue fever, and Culex pipiens, which is the dominant species of mosquitoes in the Kingdom of Saudi Arabia, as a safe method for its control. The cumulative death rate during larval development into pupae and adults was used as a criterion for evaluating tested seaweed extracts against Ae. aegypti, Cx. Pipiens. According to the obtained IC50 values ​​(the concentration that inhibits the exit of 50 % of adult mosquitoes), the results showed that C. rotundata extract (70.78 & 77.47 ppm) was more effective against A. aegypti and Cx. pipiens in comparison with H. ovata (86,98 & 95,87 ppm) and T. hemprichii (83,94 & 88,82) extracts by (1.186, 1.229, 1.146 & 1.237) fold, respectively. The results showed that the treatment with marine plant extracts against mosquito larvae of Cx. Pipiens and Ae. Aegypti gave different biological effects similar to those of other insect growth regulators (IGRs). The results also revealed the presence of morphological abnormalities in larvae that were treated with all seaweed extracts and these effects extended to all stages of growth, which caused damage to the insect without completing its life cycle. Generally, the results indicate the importance of carrying out bio-assessment tests for the pesticides that are used against mosquitoes and establishing a database to be referenced when planning control programs and making the right decision about the pesticide used.

Antioxidant, Anti-Proliferative Activity and Chemical Fingerprinting of Centaurea calcitrapa against Breast Cancer Cells and Molecular Docking of Caspase-3.

Centaurea calcitrapa has been intensively utilized in ethnomedicinal practices as a natural therapeutic recipe to cure various ailments. The current study aimed to chemically characterize ethanolic extract of C. calcitrapa (EECC) aerial parts (leaves and shoots) by use of gas chromatography-mass spectrometry analyses (GC-MS) and investigate its antioxidant and in vitro anticancer activities, elucidating the underlying molecular mechanism by use of flow cytometry-based fluorescence-activated cell sorting (FACS) and conducting in silico assessment of binding inhibitory activities of EECC major compounds docked to caspase-3. CG-MS profiling of EECC identified a total of 26 major flavonoids and polyphenolic compounds. DPPH and ABTS assays revealed that EECC exhibits potent antioxidant activity comparable to standard reducing agents. Results of the proliferation assay revealed that EECC exhibit potent, dose-dependent cytotoxic activities against triple-positive (MCF-7) and triple-negative (MDA-MB-231) breast cancer cell models, with IC50 values of 1.3 × 102 and 8.7 × 101 µg/mL, respectively. The observed cytotoxic effect was specific to studied cancer cells since EECC exhibited minimal (~<10%) cytotoxicity against MCF-12, a normal breast cell line. FACS analysis employing annexin V-FITC/propidium iodide double labeling demonstrated that the observed anti-proliferative activity against MCF-7 and MDA-MB-231 was mediated via apoptotic as well as necrotic signaling transduction processes. The increase in fluorescence intensity associated with DCFH oxidation to DCF, as reported by FACS, indicated that apoptosis is caused by generation of ROS. The use of caspase-3-specific fluorogenic substrate revealed a dose-dependent elevation in caspase-3 substrate-cleavage activity, which further supports EECC-mediated apoptosis in MCF-7 cells. The major EECC compounds were examined for their inhibitory activity against caspase-3 receptor (1HD2) using molecular docking. Three compounds exhibited the highest glide score energy of −5.156, −4.691 and −4.551 kcal/mol, respectively. Phenol, 2,6-dimethoxy established strong binding in caspase-3 receptor of hydrogenic type, with residue ARG 207 and of PI-PI stacking type with residue HIS 121. By contract, hexadecenoic acid showed 3 H-bond with the following residues: ASN 615, ASN 616a and THR 646. Taken together, the current findings reveal that EECC exhibits significant and specific cytotoxicity against breast cancer cells mediated by the generation of ROS and culminating into necrosis and apoptosis. Further investigations of the phytoconstituents-rich C. calcitrapa are therefore warranted against breast as well as other human cancer cell models.

Heavy metal ATPase genes (HMAs) expression induced by endophytic bacteria, "AI001, and AI002" mediate cadmium translocation and phytoremediation.

Contamination of heavy metals is a serious threat, which causes threats to the environment. Our study aimed to determine the role of endophytic bacteria in Cd phytoremediation and heavy metal ATPase gene expression. Cadmium (Cd) resistant endophytic bacteria were isolated from Solanum nigrum on LB agar plates, contaminated with 0-30 mg/L Cd. The phosphate solubilization and indole-3-acetic acid (IAA) production of endophytes were estimated by growing them on Pikovskaya agar medium and GC-MS analysis, respectively. An experiment in a pot was performed to evaluate the effects of bacteria on rice plants contaminated with 5-25 mg/L of Cd. Expression of Cd response genes was quantified through qRT-PCR and Cd translocation from one part to another part of the plant was measured through the ICP. BLAST alignment of 16 S-rDNA gene sequences confirmed the bacterial isolates as Serratia sp. AI001 and Klebsiella sp. Strain AI002. Both strains tolerated Cd up to 25 mg/L and produced 27-30 µg/mL of IAA. Inoculation of AI001 and AI002 improved plant growth dynamics (i.e., plant length, biomass, chlorophyll contents), relieved electrolyte leakage, and improved reduced glutathione significantly (P < 0.05). The inoculation of AI001 and AI002 significantly (P < 0.05) induced the expression of heavy metal ATPase genes ie., "HMA2, HMA3, and HMA4" and Cd translocation compared to uninoculated plants. Both AI001 and AI002 exhibited very prominent plant-growth-promoting and Cd phytoremediation properties. The results revealed that isolates also contributed a lot to the expression of rice plant heavy metal ATPase genes and in the Cd translocation in the plant.

Antioxidant enzyme responses and metabolite functioning of Pisum sativum L. to sewage sludge in arid and semi-arid environments.

The productivity of plants is a direct variant of the countless biotic and abiotic stresses to which a plant is exposed in an environment. This study aimed to investigate the capabilities of leguminous plant garden pea (Pisum sativum L.) to resist water deficit conditions in arid and semi-arid areas when applied with varied doses of sludge for growth response. The effect of sludge doses was evaluated on crop yield, antioxidant enzymes, viz., ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), superoxide dismutase (SOD), and glutathione reductase (GR), and metabolites (ascorbic acid, glutathione, and total protein content). The effective sludge concentrations with respect to seed weight and crop yield were found to be in the following trend: D2 (6.25%)>D3 (12.5%)>D1 (2.5%)>D0 (control) under organic amendment (OA). Conversely, a high dose of the sludge reduced the seed weight and total crop yield. The sludge doses D2 under arid and semi-arid conditions along with organic amendments (OA) significantly enhance the antioxidant enzyme activity, whereas sludge dose D3 with OA ominously regulates the activity of these enzymes. Besides, seeds depicted a considerable increase in ascorbic acid, glutathione, and total protein content in arid and semi-arid conditions upon the application of sludge with OA. Sewage sludge as a source of nutrients indirectly enhances crop yield, antioxidant enzymes, and antioxidant metabolites. Thus, it improves the defense mechanism, reduces abnormal protein glycation, and depletes the susceptibility of protein to proteolysis.

Analysis of phoretic relation between chewing lice and hippoboscid flies of Columba livia.

Phoresy is a biologically mechanical phenomena where an immobile organism hitches on a other mobile organism to translocate. This behaviour is not studied very well on the field level especially between two important parasites infesting the same host. Parasite/parasite interaction is rarely studied through most biological host-parasite systems. Here, we evaluated the phoretic relation between parasitic chewing lice and hippoboscid flies (Pseudolynchia canariensis) on rock pigeons. A total of 69 captivated rock pigeons, Columba livia, were examined for the parasitic chewing lice and hippoboscid flies in Giza Zoo and two local farms near Cairo, Egypt. Results indicated that there is a positive correlation between infestation of hippoboscid flies and chewing lice. Also, the analysis of louse/louse interaction using ANOVA indicated a significant difference between the three chewing louse species which were recorded on the rock pigeons with relatively high abundance of two species, Columbicola columbae and Campanulotes compar. The analysis of hippoboscid flies' abundance and its relation with chewing lice infestation indicate a significant increase of lice intensity in case of high infestation with the fly. The level of hygiene of nest may be inversely related to level of parasite infestation. This work forms a step in the process of understanding parasite/parasite and host/parasite interactions using two parasitic species with a characteristic phoretic behaviour in nature.

Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves Salix safsaf.

Organic fractions and extracts of willow (Salix safsaf) leaves, produced by sequential solvent extraction as well as infusion and decoction, exhibited anticancer potencies in four cancerous cell lines, including breast (MCF-7), colorectal (HCT-116), cervical (HeLa) and liver (HepG2). Results of the MTT assay revealed that chloroform (CHCl3) and ethyl acetate (EtOAc)-soluble fractions exhibited specific anticancer activities as marginal toxicities were observed against two non-cancerous control cell lines (BJ-1 and MCF-12). Ultra-high-resolution mass spectrometry Q-Exactive™ HF Hybrid Quadrupole-Orbitrap™ coupled with liquid chromatography (UHPLC) indicated that both extracts are enriched in features belonging to major phenolic and purine derivatives. Fluorescence-activated cell sorter analysis (FACS), employing annexin V-FITC/PI double staining indicated that the observed cytotoxic potency was mediated via apoptosis. FACS analysis, monitoring the increase in fluorescence signal, associated with oxidation of DCFH to DCF, indicated that the mechanism of apoptosis is independent of reactive oxygen species (ROS). Results of immunoblotting and RT-qPCR assays showed that treatment with organic fractions under investigation resulted in significant up-regulation of pro-apoptotic protein and mRNA markers for Caspase-3, p53 and Bax, whereas it resulted in a significant reduction in amounts of both protein and mRNA of the anti-apoptotic marker Bcl-2. FACS analysis also indicated that pre-treatment and co-treatment of human amniotic epithelial (WISH) cells exposed to the ROS H2O2 with EtOAc fraction provide a cytoprotective and antioxidant capacity against generated oxidative stress. In conclusion, our findings highlight the importance of natural phenolic and flavonoid compounds with unparalleled and unique antioxidant and anticancer properties.

Highly Water-Soluble Luminescent Silica-Coated Cerium Fluoride Nanoparticles Synthesis, Characterizations, and In Vitro Evaluation of Possible Cytotoxicity.

A coprecipitation process was utilized for the preparation of terbium fluoride nanocrystals by cerium fluoride. Silica was used to modify the surface of these core/shell nanocrystals. The synthesized CeF3:Tb@LaF3 and CeF3:Tb@LaF3@SiO2 nanoparticles (NPs) were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), UV/vis spectrophotometry, and photoluminescence spectrophotometry. XRD patterns showed resolved reflection planes with broad widths, confirming the nanocrystalline nature of the CeF3:Tb@LaF3@SiO2 NPs. Fourier transform infrared spectra clearly revealed a uniform, smooth silica layer encapsulating the luminescent seed core and confirmed the polycrystalline nature of the CeF3:Tb@LaF3@SiO2 NPs. The TEM result showed an average crystalline size of 18 nm, which illustrated good agreement with the XRD results. The results of photoluminescence spectrophotometry confirmed the doping of terbium ions in the CeF3 crystal lattice. The cytotoxicity results of the MTT assay showed that CeF3:Tb@LaF3@SiO2 NPs have minimum toxicity with respect to CeF3:Tb@LaF3 NPs and the control drug dasatinib on HT-29 and HepG2 cell lines. Moreover, results of inverted microscopy confirmed the nontoxic and biocompatible nature of CeF3:Tb@LaF3@SiO2 NPs. These findings show that CeF3:Tb@LaF3@SiO2 NPs are promising candidates for applications in biomedical science in the future, such as bioimaging, biolabeling, biodetection or bio-probing, labeling of cells and tissue, drug delivery, cancer therapy, and multiplexed analysis.

Effects of various doses of glutathione on the proliferation, viability, migration, and ultrastructure of cultured human melanocytes.

Normal human cultured melanocytes were exposed to various glutathione concentrations (0.1, 0.5, 1.0, 5.0, and 10.0 mg/mL) for 72 hours. At the end of the experiment, proliferation, viability, migration, and ultrastructural changes were monitored. Glutathione at the doses of 0.5 to 10.0 mg/mL reduced the viability of melanocytes significantly as compared to the control (P < .05). Glutathione significantly reduced the proliferation of melanocytes at the doses of 0.5 to 10.0 mg/mL as compared to the control (P < .001). Glutathione at 0.5 to 10.0 mg/mL significantly reduced the migration of melanocytes as compared to the control (P < .001). The percentage of mature melanosomes was 53.43% in control and 50.58%, 41.83%, 33.4%, and 8.95% in 0.1, 0.5, 1.0, and 5.0 mg/mL glutathione exposed cells, respectively. This reduction in the number of mature melanosomes was statistically significant as compared to the control. However, no cytotoxic effects were recognized by electron micrographs. These results encourage the potential implementation of glutathione as a skin-lightening agent. However, this study is limited by cell culture and ultrastructural. It should therefore be expanded in the future to include patients with pigmentary disorders.

Safety and efficacy of parenteral glutathione as a promising skin lightening agent: A controlled assessor blinded pharmacohistologic and ultrastructural study in an animal model.

Hyperpigmentation was induced in the skin of experimental animals using UVB at 6 J/cm2 three times a week for three consecutive weeks. Subsequently, glutathione was injected intraperitoneally in the experimental animals at doses of 10, 20, and 40 mg/kg body weight three times a week for three consecutive weeks. At the end of the experiment, blood samples and lung, kidney, liver, and skin tissue specimens were collected from animals for hematological, biochemical, histological, and electron microscopy examination. Glutathione at 40 mg/kg body weight/day reduced skin hyperpigmentation significantly, except at low doses. The skin lightening effect assessed by a chromameter was dose-dependent. There were no statistically significant differences among the mean values of AST, ALT, creatinine, BUN, and CBC counts across the four groups. Lung, kidney, and liver tissue specimens did not show any histological toxic changes. The number of melanin granules was significantly lower in the group treated with the highest dose of glutathione compared to that in the control. Electron microscopy proved that glutathione at 20 and 40 mg/kg body weight/day was able to reduce the number of melanized cells significantly compared to that in the control. Parenteral glutathione was effective as a skin lightening agent and did not provoke any toxic effects in the employed animal model. The limitation of the study was conducted in guinea pigs and was of short-term duration.

Nano-erythrocyte membrane-chaperoned 5-fluorouracil liposomes as biomimetic delivery platforms to target hepatocellular carcinoma cell lines.

Nano-erythrocyte coating has been developed as an interesting biomimetic platform to provide hybrid nano-carriers with innate functions to target liver cancer. This goal was achieved by coating nano-erythrocyte membranes (NEMs) onto 5-fluorouracil (5-FU)-loaded liposomes (LPs) to produce NEM-5-FU-LPs. This framework is used to promote the escape of 5-FU-LPs from degradation during systemic circulation. NEMs were obtained by hypotonic lysis of erythrocytes to produce ghost erythrocytes (GEs) followed by extrusion through polycarbonate membranes. Chimeric NEM-5-FU-LPs were fabricated via the fusion of NEMs and artificial LPs. The resultant chaperoned LPs were characterized based on particle size, morphology, entrapment efficiency (EE %), stability, protein content and phosphatidylserine exposure and their in vitro release profiles and cytotoxic efficacy were also determined. The present results revealed that 5-FU-LPs, NEM-5-FU and NEM-5-FU-LPs exhibited nanosize, spherical shapes and unimodal size distributions <0.3. In addition, the vesicles presented a zeta potential with EE% of 24.6-30.7% and an appropriate stability for 3 weeks. NEM-5-FU-LPs retained the erythrocyte membrane proteins as confirmed by PAGE and displayed a sustained release profile up to 48 h when compared to NEM-5-FU and the 5-FU solution. Moreover, hybrid NEM-5-FU-LPs induced a late cytotoxic effect after 48 h compared to the other formulations. Thus, mantling of 5-FU-LPs by NEMs could enhance vesicle controllability and their targetability to liver cancer cells.